Electroantennogram responses of the Triatoma dimidiata complex to volatiles produced by its exocrine glands.

Members of the Triatoma dimidiata complex are vectors of the protozoan parasite Trypanosoma cruzi, the etiologic agent of Chagas disease. Morphological and genetic studies indicate that T. dimidiata complex has three principal haplogroups in Mexico. However, whether there are differences in the olfactory physiology among the haplogroups of this complex and a possible correlation with their antennal phenotype are not yet known. Antennal responses to 13 compounds released from the metasternal and Brindley´s glands, which are involved in the alarm and mating-related behaviours of T. dimidiata were investigated using electroantennography (EAG). Overall, of the 13 compounds tested, seven triggered EAG responses in both sexes of three Mexican haplogroups. The sensitivity of the EAG responses show some relationship with the total number of chemo-sensilla present on the antennae. Antennal sensitivity was different between sexes and haplogroups of the T. dimidiata complex. Discriminant analysis of EAG sensitivity was significant, separating the three haplogroups. Our finding is consistent with morphological and genetic evidence for haplogroups distinction within the complex.

A CO2-Free Synthetic Host-Odor Mixture That Attracts and Captures Triatomines: Effect of Emitted Odorant Ratios.

Triatomines, vectors of Chagas Disease, are hematophagous insects. Efforts have been made to develop synthetic attractants based on vertebrate odor-to lure them into traps. However, because those lures are not practical or have low capture efficiency, they are not in use in control programs. Therefore, more work is needed to reach a practical and efficient odor lure. Recently, a three-component, CO2-free, synthetic blend of vertebrate odor (consisting of ammonia, l-(+)-lactic acid, and hexanoic acid), known as Sweetscent (Biogents AG, Regensburg, Germany), was shown to attract and capture triatomines in the laboratory. In this study, using a trap olfactometer and an odor blend with constituents similar to those of Sweetscent (delivered from low-density polyethylene sachets) we found that the odorant ratios of the mixtures have a strong effect in the capture of triatomines. The blend with the most efficient combination of odorant ratios evoked ca. 81% capture in two relevant triatomine species. In the case of the most effective odor mixtures, we measured the odor mass emission for the three components of the mixture and therefore were able to estimate the odorant ratios emitted that were responsible for such a high capture performance. Thus, in those mixtures, pentanoic acid was the main component (ca. 65 %) followed by ammonia (ca. 28%) and, l(+)-lactic acid (ca. 7 %). Our results are encouraging as efficient, practical, and cheap odor baits to trap triatomines in the field would be within reach. The odor-delivery system used should be improved to increase stability of odor emission.

Morphology and physiology of the olfactory system of blood-feeding insects.

Several blood-feeding (hematophagous) insects are vectors of a number of diseases including dengue, Chagas disease and leishmaniasis which persistently affect public health throughout Latin America. The vectors of those diseases include mosquitoes, triatomine bugs and sandflies. As vector control is an efficient way to prevent these illnesses it is important to understand the sensory biology of those harmful insects. We study the physiology of the olfactory system of those insects and apply that knowledge on the development of methods to manipulate their behavior. Here we review some of the latest information on insect olfaction with emphasis on hematophagous insects. The insect olfactory sensory neurons are housed inside hair-like organs called sensilla which are mainly distributed on the antenna and mouthparts. The identity of many of the odor compounds that those neurons detect are already known in hematophagous insects. They include several constituents of host (vertebrate) odor, sex, aggregation and alarm pheromones, and compounds related to egg-deposition behavior. Recent work has contributed significant knowledge on how odor information is processed in the insect first odor-processing center in the brain, the antennal lobe. The quality, quantity, and temporal features of the odor stimuli are encoded by the neural networks of the antennal lobe. Information regarding odor mixtures is also encoded. While natural mixtures evoke strong responses, synthetic mixtures that deviate from their natural counterparts in terms of key constituents or proportions of those constituents evoke weaker responses. The processing of olfactory information is largely unexplored in hematophagous insects. However, many aspects of their olfactory behavior are known. As in other insects, responses to relevant single odor compounds are weak while natural mixtures evoke strong responses. Future challenges include studying how information about odor mixtures is processed in their brain. This could help develop highly attractive synthetic odor blends to lure them into traps.

Evaluation of a CO2 -free commercial mosquito attractant to capture triatomines in the laboratory.

Efforts have been made to develop vertebrate odor-based attractants to lure hematophagous triatomines into traps. However, more work is needed to reach a practical, cheap, and efficient odor lure. We carried out attraction and capture tests in a dual-choice olfactometer and a pitfall trap. Here we report that a three-component, CO2 -free, synthetic blend of vertebrate odor (consisting of ammonia, L(+) lactic acid and hexanoic acid, and known as Sweetscent®) significantly induces 3(rd) -instar Rhodnius prolixus and Triatoma infestans nymphs to fall into the test capture-tube of the olfactometer. Blend constituents presented singly or in two-component blends did not evoke a response and, therefore, we propose that the insects respond specifically to the three-component blend in a synergistic way. When tested in a pitfall trap in an experimental arena, this blend induced capture in 37.5% of the lured traps, whereas 9% of the nymphs tested were captured in a single night. No insects were captured in control traps. Our work represents a proof-of-concept regarding capture of triatomines using host odor-based, CO2 -free synthetic mixtures as lures for pitfall traps. CO2 -free lures are more practical for field work than natural or CO2 -containing synthetic blends.

Olfactory and behavioural responses of the blood-sucking bug Triatoma infestans to odours of vertebrate hosts.

Olfactory receptors in basiconic and grooved-peg sensilla on the antenna of fifth-instar Triatoma infestans nymphs respond to host odours. Gas chromatography analyses of host odour extracts coupled to electrophysiological recordings from basiconic sensillum receptors indicate that nonanal is a constituent of sheep wool and chicken feather odour that stimulates one of the receptors in this type of sensillum. Similar analyses revealed isobutyric acid in rabbit odour to be a chemostimulant for one of the receptors in grooved-peg sensilla. The response of the aldehyde receptor was higher to heptanal, octanal and nonanal than to other aliphatic aldehydes, and the response of the acid receptor was higher to isobutyric acid than to other short-chain branched and unbranched acids. The behavioural responses of fifth-instar T. infestans nymphs to nonanal and isobutyric acid in an air-stream on a servosphere indicate that, whereas nonanal causes activation of the bugs, isobutyric acid induces an increase in upwind displacement, i.e. odour-conditioned anemotaxis. Binary mixtures of these compounds did not improve the attraction obtained with isobutyric acid alone. A comparison of the behavioural and electrophysiological responses of the bugs to different amounts of isobutyric acid in air suggests that attraction is obtained at concentrations that causes low-to-moderate increases in the firing rate of the acid-excited receptor in the grooved-peg sensilla, whereas at a dose that evokes relatively high firing rates (>40 Hz) no attraction is obtained.

Baker's yeast, an attractant for baiting traps for Chagas' disease vectors.

We tested the attraction of volatile compounds, produced by the aerobic growth of Saccharomyces cerevisiae on saccharose for Triatoma infestans. For these tests, we exploited the behavioural characteristic of these haematophagous insects of dropping when searching for food. In olfactometer assays, yeast cultures activated and attracted bugs as effectively as a mouse. The attraction of the cultures was significantly reduced when the carbon dioxide released was partially eliminated using potassium hydroxide. Yeast cultures were also tested as lures in a novel trap device. A baited device for trapping Chagas' disease vectors using the behavioural peculiarities of T. infestans and this simple attractant is described.